CN109646561A

CN109646561A - A.C.C. extract with anti-inflammatory and antibacterial effect and the composition comprising it as effective component

Info

Publication number: CN109646561A
Application number: CN201811072025.7A
Authority: CN
Inventors: 禹勇奎
Original assignee: Woo Young Gyo
Current assignee: Woo Young Gyo
Priority date: 2017-12-28
Filing date: 2018-09-14
Publication date: 2019-04-19
Also published as: US11590191B2; CN112805021A; US20200353028A1; DE112018006698T5; WO2019132147A1; KR20190079849A; KR102118540B1

Abstract

The present invention is provided by Cortex Phellodendri (Phellodendron bark), radix scutellariae (Scutellaria baicalensis), Chinese herbaceous peony (Paeonia lactiflora Pall), shaggy-fruited dittany (Dictamnus dasycarpus Turcz), rhizoma anemarrhenae (Anemarrhena asphodeloides), alum (Alumen), kapur (Dryobalanops aromatica Gaertner), cornmint (Mentha arvensis var.Piperascens), elecampane (Inula helenium), Korea's open country fourth A.C.C. extract and the composition comprising it as effective component made of fragrant (Syringa velutina var.Kamibayashii), C. incisa (Corydalis incisa), Eclipta prostrata (Eclipta prostrata), honeysuckle (Lonicera japonica) and the cooperation of Radix Glycyrrhizae (Glycyrrhiza uralensis) Chinese medicine.A.C.C. extract of the invention, which is presented, reduces tumor necrosis factor-alpha (tumor necrosis factor- α, TNF-α), interleukin-1 ' beta ' (interleukin-1 β, IL-1 β) and interleukin-6 (IL-6) generation, the anti-inflammatory effect for adjusting the expression of iNOS NO is inhibited to generate, it is a kind of substance for clinically alleviating the symptoms such as diaper rash and prickly heat also by anti-inflammatory and antibacterial action, can be used as effective inflammation treatment agent.

Description

A.C.C. extract with anti-inflammatory and antibacterial effect and comprising its as effectively at The composition divided

Technical field

The present invention relates to extracts and antibacterial and anti-inflammatory composition with anti-inflammatory and antibacterial effect.

Background technique

Restore impaired portion when inflammatory reaction caused by vivo is physics, the chemical stimulation by scar or bacterium infection etc One of the important body defenses mechanism of position, when being stimulated, vaso-active substance local ionization, thus blood vessel permeability increases Add, induces inflammation.Also, inflammatory reaction is the recovery system formed in vivo according to outside stimulus, clinically along with Breaking-out, fever, swelling, pain etc. and occur.The relevant cell of inflammatory reaction has macrophage, single glomus cell, lymph corpuscle, hypertrophy Cell, fiber sprout cell, blood platelet etc., macrophage (machr ophage) are referred to as causing the main cell of inflammatory reaction, These are activated the case where being stimulated or by the cell factor (cytokine) of immunocyte secretion, generate nitric oxide (nitric oxide) (NO) and cytokine, play an important role to biophylaxis.Also, when inflammatory reaction, generates and release Put the inflammation of histamine, thrombocytin, arachidonic acid (arachidonic acid) metabolism group, active oxygen, cell factor or the like Disease mediating factors.

Inflammatory reaction repeatedly causes the excessive generation of reactive oxygen species and active nitrogen cluster in inflammatory cell, causes permanent Thus transgenosis can also result in irreversible lesion.Although that is, inflammatory reaction be in vivo caused by defense mechanism, When persistently causing inflammatory reaction, promotes mucosa injury, finally cause edema, pain, breaking-out, fever etc., can lead to cardiovascular disease The disease relevant to chronic inflammation of disease, rheumatoid arthritis, bronchitis and cancer etc.

Taking it by and large, the iNOS and inflammatory reaction phase in nitricoxide synthase (nitric oxide synthase, NOS) It closes, is expressed because of the stimulation of gamma interferon (interferon- γ), inflammatory cytokine and LPS.It is well known that intracorporal NO takes on critical function in antibacterial and tumor resection, but causes increase, the edema etc. of blood vessel permeability by the NO that iNOS is formed Reaction, promotes the synthesis of inflammation mediator, makes increased inflammation.

It is well known that lipopolysaccharides (Lipopolysaccharide, LPS) is the endogenous toxic material in the epicyte of gram-negative bacteria Element, this stimulating expression of macrophage or single glomus cell, stimulate tumor necrosis factor-alpha (tumor necrosis factor- α, TNF- α), the inflammatory mediators cell such as interleukin-1 ' beta ' (interleukin-1 β, IL-1 β) and interleukin-6 (IL-6) because The secretion of sub (cytokine), Lai Cuihua inflammatory reaction.TNF-α is that middle macrophage, lymph corpuscle and leucocyte etc. are raw in vivo At cell factor (cytokine), do not generate in normal state, if macrophage is stimulated, is synthesized and secretes. IL-6 is the one kind of representative pro-inflammatory cytokine (pro-inflammatory cytokine), from including single glomus cell Various kinds of cell in secrete, in the early stage immune response in play an important role.It is well known that IL-1 β is together with IL-6 and TNF-α It indicates one of representative inflammatory cytokine, is the medium substance of generation NO, and be to cause local inflammation, and realize T The cytokine of the maturation and NK cell of cell-stimulating and B cell activation.It is reported that the synthesis of this inflammatory mediator substance With by Cycloxygenase (cyclooxygenase, COX), arachidonic acid (arachidonic acid) is converted into leukotriene (leukotriene), the process and nitric of prostaglandin (prostaglandin), thromboxane (thromboxane) etc. The generation of oxide (NO) is related, finally brings mortality result to host.Especially, NO is substance with high reactivity, and is The substance of effect needed for executing innate immune response to the pathogen of bacterium, virus, bacterium and helminth etc, passes through NO Synthase (NOS) is made by L-arginine (L-arginine), NOS point for primary type (constitutive NOS, CNOS it) and induces type (inducible NOS, iNOS).It is known that iNOS is when by outside stimulus or pro- When the stimulation of inflammatory cytokine etc., a large amount of NO is produced, causes blood vessel permeability and edema etc., induces inflammation It reacts, under serious situation, also spreads to the immunity disease of rheumatoid arthritis and autoimmune disease etc.With this phase Guan Di needs to develop the Nature inorganic bone object with antibacterial and anti-inflammatory activity and effective anti-inflammatory therapy agent together.

Summary of the invention

The present invention, which provides, utilizes natural goods Chinese medicine, and the extract with antibacterial and antiphlogistic effects, especially, purpose exists In confirming that the anti-inflammatory of A.C.C. extract for carrying out distillation extraction to total 14 kinds of Chinese medicine comprising Cortex Phellodendri and obtaining is living Property, and confirm clinical effectiveness.

Another object of the present invention is to confirmation A.C.C. extract is that there is anti-inflammatory effect and potent antibacterial to imitate together The effective component of fruit.

Also, it is intended that confirmation A.C.C. extract is clinically also slow by anti-inflammatory and antibacterial action The substance for solving the symptom of diaper rash and prickly heat etc., thereby confirms that A.C.C. extract can be used as effectively treating diseases associated with inflammation Agent.

The present invention to solve the above-mentioned problems, is provided by Cortex Phellodendri (Phellodendron bark), radix scutellariae (Scutellaria baicalensis), Chinese herbaceous peony (Paeonia lactiflora Pall), shaggy-fruited dittany (Dictamnus Dasycarpus Turcz), rhizoma anemarrhenae (Anemarrhena asphodeloides), alum (Alumen), kapur (Dryobalanops aromatica Gaermer), cornmint (Mentha arvensis var.Piperascens), soil Radix aucklandiae (Inula helenium), Korea open country cloves (Syringa velutina var.Kamibayashii), C. incisa (Corydalis incisa), Eclipta prostrata (Eclipta pro strata), honeysuckle (Lonicera japonica) and Radix Glycyrrhizae A.C.C. extract made of the cooperation of (Glycyrrhiza uralensis) Chinese medicine.

Especially, which is characterized in that the formula for being used to prepare the Chinese medicine of said extracted object is respectively Cortex Phellodendri (Phellodendron bark) 16~24 weight percent, 1~7 weight hundred of radix scutellariae (Scutellaria baicalensis) Divide ratio, Chinese herbaceous peony (Paeonia lactiflora Pall) 5~11 weight percent, shaggy-fruited dittany (Dictamnus dasycarpus Turcz) 5~11 weight percent, rhizoma anemarrhenae (Anemarrhena asphodeloides) 1~7 weight percent, alum (Alumen) 4~12 weight percent, kapur (Dryobalanops aromatica Gaertner) 1~7 weight percent Than, cornmint (Mentha arvensis var.Piperascens) 1~7 weight percent, elecampane (Inula Helenium) 1~7 weight percent, Korea open country cloves (Syringa velutina var.Kamibayashii) 1~7 weight Percentage, C. incisa (Corydalis incisa) 1~7 weight percent, Eclipta prostrata (Eclipta prostrata) 1~7 weight Amount percentage, honeysuckle (Lonicera japonica) 1~7 weight percent, Radix Glycyrrhizae (Glycyrrhiza uralensis) 1~ 7 weight percent.

Said extracted object, which is characterized in that total 14 kinds of Chinese medicine comprising Phellodendron bark (Cortex Phellodendri) The sample (A.C.C. extract) for carrying out distillation extraction and obtaining has antibacterial and anti-inflammatory activity.In related experiment, confirmation The nitric oxide of cell is discharged in 264.7 cell of RAW stimulated by lipopolysaccharides (Lipopolysaccharide, LPS) (NO) production quantity and inflammatory cells tumor necrosis factor-alpha (tumor necrosis factor- α, TNF-α), leucocyte are situated between The variation of -1 β of element (interleukin-1 β, IL-1 β) and interleukin-6 (IL-6) production quantity.As a result, A.C.C is extracted Object no cytotoxicity it is potent inhibition by the generation of LPS increased NO and inflammatory cytokine.

And, which is characterized in that A.C.C. extract of the invention is to Pseudomonas aeruginosa (Pseud omonas Aeruginosa), staphylococcus aureus (Staphylococcus aureus), methicillin-resistant staphylococcus aureus (MRSA, Methicillin-resistant Staphylococcus aureus), candida albicans (Candida albicans) And variation hammer (Streptococcus mutans) bacterium has antibacterial action.A.C.C. extract is presented in above-mentioned bacterium 99.9% potent bacterial reduction, it means that A.C.C. extract is having with anti-inflammatory effect and antibacterial effect together Imitate ingredient.

In related experiment, Pseudomonas aeruginosa (Pseudomonas aeruginosa), staphylococcus aureus are utilized (Staphylococcus aureus), methicillin-resistant staphylococcus aureus (MRSA, Methicillin-resistant Staphylococcus aureus), candida albicans (Candida albicans) and variation hammer (Streptococcus Mutans) bacterial strain confirms antibacterial activity, its clinic is applicable in, to the infant for suffering from diaper rash, pruritus, prickly heat symptom When being coated with A.C.C. extract, eruption is presented, allergic skin, festers, the symptoms improvement such as pruritus, prickly heat, meat can be used The prickly heat of eye confirmation infant's skin is alleviated pretty good after 2 weeks.Thus A.C.C. extract can be confirmed in clinic Upper is also the substance for alleviating the symptoms such as diaper rash and prickly heat by anti-inflammatory and antibacterial action, and A.C.C. extract can become pair The effective alternative solution of diseases associated with inflammation.

Another aspect of the present invention, which is characterized in that provide comprising above-mentioned A.C.C. extract as the anti-of effective component Bacterium and anti-inflammatory composition.Since the synergistic effect of anti-inflammatory effect and antibacterial action is presented in above-mentioned A.C.C. extract, It can be used as effective inflammation treatment agent.

The A.C.C. extract for carrying out distillation extraction to a total of 14 kinds of Chinese medicine comprising Cortex Phellodendri and obtaining has together There are anti-inflammatory effect and potent antibacterial effect.

Also, A.C.C. extract of the invention reduces the generation of TNF-α, IL-1 β and IL-6, is thus presented by adjusting The expression of iNOS and inhibit NO generate anti-inflammatory effect.

Therefore, A.C.C. extract of the invention is clinically also to alleviate diaper rash by anti-inflammatory and antibacterial action And the substance of the symptoms such as prickly heat, it can be used as effective inflammation treatment agent.

Detailed description of the invention

Fig. 1 is the schematic diagram that the distillation of 14 natural goods comprising phellodendron bark (Cortex Phellodendri) is extracted.

Fig. 2 is to indicate using the confirmation of MTT assay method with LPS processing A.C.C extract and to 264.7 cell band of Raw The chart of the result for the toxicity come.(*, p < 0.05, * *, p < 0.01compared to control.Con；Control, LPS 1 μg/ml；1 μ g/mllipopolysaccharide, A0；LPS 1 μ g/ml, A1；A.C.C.1 μ g/ml, A5；A.C.C.5 μ g/ml, A10；10 μM of A.C.C.10 μ g/ml, Q.C；quercetin 10μM.)

Fig. 3 is to indicate to be induced in 264.7 cell of Raw by LPS and increased NO generates concentration due to A.C.C. extract The chart inhibited to dependence.(###, p < 0.001compared to control, and***, p < 0.001compared to 1μg/ml LPS.Con；1 μ g/ml of control, LPS；1 μ g/ml lipopolysaccharide, A0；LPS 1 μ g/ml, A1； A.C.C.1 μ g/ml, A5；A.C.C.5 μ g/ml, A10；10 μM of A.C.C.10 μ g/ml, Q.C；quercetin 10μM.)

Fig. 4 is to indicate that A.C.C. extract generates the figure for inhibiting bring effect to the cytokine of 264.7 cell of Raw Table.

(a) A.C.C. extract is on dose-dependent influence (the Effects of A.C.C.extracts on of TNF-α TNF-αin dose-dependent manner)。

(b) A.C.C. extract is on dose-dependent influence (the Effects of A.C.C.extracts on of IL-1 β IL-1β in dose-dependent manner)。

(c) A.C.C. extract is on dose-dependent influence (the Effects of A.C.C.extracts on of IL-6 IL-6in dose-dependent manner)。

Pro-inflammatory cytokines is the Raw264.7 cell stimulated by LPS, is surveyed in the medium It is fixed.1 μ of ###, p < 0.001compared to control, and**, p < 0.01, and***, p < 0.001compared to g/ml LPS.Con；1 μ g/ml of control, LPS；1 μ g/ml lipopolysaccharide, A0；LPS 1 μ g/ml, A1； A.C.C.1 μ g/ml, A5；A.C.C.5 μ g/ml, A10；A.C.C.10μg/ml.

Fig. 5 is the chart for indicating the symptom variation using diaper rash after A.C.C. extract and prickly heat etc..(n= 18) .*, p < 0.05, and***, p < 0.001compared to 0week

Fig. 6 is the picture for indicating the symptom variation using diaper rash after A.C.C. extract and prickly heat etc..

Specific embodiment

Hereinafter, by embodiment, the more detailed description present invention.But these embodiments only example of the invention The property shown contents, and the scope of the invention is not limited to these embodiments.

1. material of embodiment select and the preparation of A.C.C. extract

Based on Herbal Textuals, file investigation is carried out, picks out 14 kinds of Chinese medicine in total.Chinese medicine has purchased to be enabled by Soul medicine The product for being packaged as crude drug preparation and selling that the CK (strain) in city produces.Dahl Burke Improved Eagle Medium (Dulbecco ' S modified eagle ' s me dium, DMEM), fetal calf serum (fetal bovine serum, FBS) purchase in Hai Kelong (Hyclone) (Logan, UT, USA).Penicillin (Penicillin) (100U/mL), streptomysin (streptomycin) (100ug/ml) purchase in Gibco (life technology Co., Ltd, Gaithersburg, MD, USA, Life technology 1nc., Gaithersburg, MD, USA).LPS, 3- (4,5- dimethylthiazole -2- base) -2,5- diphenyltetrazolium bromide bromide (3- (4,5- Dimethylthiazol-2-y1) -2,5-diphenyl tetrazolium bromide, MTT), sodium nitrite (Sodium Nitrite), dimethyl sulfoxide (Dimethyl sulfoxide) (DMSO) buys sigma chemistry product company (Sigma Chemical Co.) (St.Louis, MO, USA).Nitrite oxide (NO) detection kit (detection Kit) purchase It is used in introne (iNtron), elisa assay kit (assaykit) is bought in Thermo Fischer Scient Inc. (Thermo Fisher Scientific)。

According to each ratio, cooperate 13 kinds of Chinese medicines other than Cortex Phellodendri, third distilled water 40L is used as solvent, 11 hours (referring to Fig.1, table 1) is extracted in distillation at 80 DEG C.

Table 1

Ratio in 1. material of table and A.C.C. extract

Experimental example 1: the cytotoxicity assay based on MTT assay

The culture of 264.7 cell of Raw

264.7 cell of Raw is bought in Korea Cell strain bank.Using contain Penicillin/stre ptomycin The DMEM culture medium of 100unit/ml and 10%FBS, the culture in 37 DEG C, 5%CO2 incubator (incubator), every 3 days one Secondary carry out squamous subculture.

96 orifice plates (well plate) are adjusted to 2 × 10 by the measurement of 264.7 cells survival rate of Raw⁴Cells/well's After concentration, cultivates 24 hours, execute stabilisation.After 24 hours, the culture medium of well is all removed.A.C.C. to be extracted The mode that the ultimate density of object reaches 1,5,10 μ g/ml is diluted in culture solution, and handles after cell strain, cultivates 3 hours.3 After hour, 1 μ g/ml of LPS is handled in the well other than control group.After removing culture medium after 24 hours, each Well adds MTT solution (0.2 μ g/ml is without phenol red DMEM, 0.2 μ g/ml in phenol red free DMEM) 100 μ l, 37 DEG C, react 2 hours in 5%CO2incubator, restore MTT.Later, it is given birth in each well addition DMSO solution to dissolve At formazan (formazan).Using microplate reader (Microplate reader), absorbance is measured at 545nm.

The measurement of experimental example 2:Nitric Oxide production quantity

In 6well plate, with 1 × 10⁵The concentration of cells/well adds 264.7 cell of Raw.In 37 DEG C, 5% After taking care of 24 hours in CO2incubator, 1,5,10 μ g/ml A.C.C. extracts are handled in 264.7 cell of Raw, 3 is small When after, handle 1 μ g/ml of LPS, cultivate 24 hours.With the nitrite for being based on Griess reaction (Griess reaction) Oxide (NO) detection kit (iNtron) analyzes nitric oxide.Ge Lisi is put into cell culture supernatant Reagent (Griess Reagent) simultaneously mixes, and after reacting 10 minutes at normal temperature, utilizes microplate reader (microplate Reader), measured under 520nm absorbance.Based on the various concentration standard curve of sodium nitrite (Sodium nitrite), fit For in formula, determining the NO concentration in culture solution.

Experimental example 3: the measurement of inflammation correlation cytokines secretory volume

1 × 10 is added in 6well plate⁵264.7 cell of Raw of the concentration of cells/well, to measure conduct TNF-α, IL-6 the and IL-1 β production quantity of inflammation induction cell factor.After 24 hours, 1,5,10 μ g/ml A.C.C. of processing are mentioned Object is taken, after 3 hours, the LPS of 1 μ g/ml is handled in all well other than control group.After culture 24 hours, benefit With ELISA kit (Thermo Fisher Scientific), TNF-α of the analysis secretion in culture medium, IL-6 and IL-1 β.Benefit With Microplate reader, measure at 450 nm.

Experimental example 4: the measurement of antibacterial activity

In order to watch the antibacterial activity of A.C.C. extract, antimicrbial power is measured using paper disc (paper disc).Test it The test strain of preceding culture, 1.0 × 10⁴It is seeded in each culture medium above.A.C.C.100 μ l is loaded in the paper disk of sterilizing Later, 30 minutes dry, with spare.On the culture medium of inoculation test bacterial strain, the paper for being mounted with A.C.C. is placed at intervals After disk, cultivated 24-48 hours in incubator.Measure the periphery Paper disk inhibition zone, carry out result judgement and Analysis.

Experimental example 5: clinical research

The subject convened by bulletin makes " preparatory questionnaire " by agent (guardian) according to Properties of Objects. By preparatory questionnaire survey, after grasping symptom, skin condition etc., composition is provided, it is same daily that researcher educates subject The time of sample is coated with 5-10 times with the amount of daily 1cc, so coating 14 days.Also, for objective result of study, shooting is suffered from Locate picture, comparative analysis coating before with coating after, at this time in picture except as face can recognize individual position.Coating 14 After it, agent's production " test questionnaire ".According to life ethics method, the time terminated from research is lighted for the keeping organ of data Keeping 3 years, during information protection after, personal information and picture will pass through irrecoverable method permanent delet.

Experimental example 6: statistical analysis

All measured values are indicated by average ± standard deviation value, examine (t-test) suitably by two sides independent sample t- Or variance analysis test (ANOVA test) executes statistical analysis.Cell mean between each group shows sizable difference When, by post method Scheffe ' s method, it is applicable in the analysis.

Experimental result

1.A.C.C. extract is to cytotoxicity caused by 264.7 cell of Raw

In order to confirm A.C.C. extract on influence caused by 264.7 cell of Raw, by various concentration (1,5,10 μ g/m1) After pre-processing A.C.C. extract, the LPS of 1 μ g/ml is handled.After culture 24 hours, MTT assay method, confirmation are utilized Cytotoxicity.The confirmation of its result, cells survival rate is respectively 98.47,90.91,93.23%, is not presented to cells survival significant The high survival rate (Fig. 2) of meaning.

2.A.C.C. extract, which generates the NO of 264.7 cell of Raw, inhibits bring effect

When with macrophage-stimulating LPS, inducible nitric oxide synthase (inducible nitric oxide is expressed Synthase, iNOS), generate excessive NO.The NO of generation promotes inflammatory reaction, induces tissue damage.Therefore, it to report A.C.C. whether extract inhibits the inflammatory mediators substance of NO etc.The confirmation of its result handles LPS in 264.7 cell of Raw When, compared with the control group, NO is generated from 0.45 and is increased to 3.15, that is, increases by 7 times.It is mentioned when with 1,5,10 μ g/ml processing A.C.C. When taking object and measuring the generation inhibitory effect of NO, result confirmation, in 264.7 cell of Raw due to LPS is induced increased NO Generation is suppressed to concentration dependent (referring to Fig. 3) due to A.C.C. extract.

3.A.C.C. extract, which generates the cytokine of 264.7 cell of Raw, inhibits bring effect

During inducing inflammatory reaction, along with the generation of the inflammatory mediator object of NO and PGE2 etc and by immune The generation of the inflammatory cytokine of reaction.Representative cytokine has TNF-α, IL-1 β and IL-6.In order to watch A.C.C. Which kind of influence extract brings to inflammatory mediator cytokine generation, by ELISAassay method, measures TNF-α, IL-1 β And the variable quantity of IL-6.By various concentration, after 264.7 cell of Raw handles A.C.C. extract, LPS processing is carried out.Its As a result, 264.7 cell of Raw handle LPS when, compared with the control group, TNF-α, IL-1 β and IL-6 increase quite a lot of (figure 4).But by handling A.C.C. extract, reduce TNF-α, IL-1 β and IL-6.Especially, it is handled when with 10 μ g/ml concentration A.C.C. when extract, reduce quite a lot of.Confirmed by this result, A.C.C. extract inhibits in 264.7 cell of Raw The cytokine of middle induction inflammation, thus has anti-inflammatory effect.

The antibacterial activity effect of 4.A.C.C. extract

As itch, the main pathogenic fungi of children's eruption, it is known that A group streptococcus (Streptococcus pyogenes) and Staphylococcus aureus (Staphylococcus aureus).Also, as women vaginitis (Vaginal Candidiasis), eruption, scratchiness main pathogen fungi, have Candida albicans (Candidaalbicans), as a result, when The skins such as the thigh root of male fold and humidity position by mycotic infection when, jock itch or intertriginous candidiasis etc can occur Skin infection disease.In this experiment, by the method for following table, to Pseudomonas aeruginosa (Pseudomonas aeruginosa), Staphylococcus aureus (Staphylococcus aureus), methicillin-resistant staphylococcus aureus (MRSA, Methicillin-resistantStaphylococcus aureus), candida albicans (Candida albicans) and variation Hammer (Streptococcus mutans) bacterium is tested, and A.C.C. extract is presented in the bacterial strain for being total up to 5 kinds 99.9% potent bacterial reduction (%) (table 2).

Table 2

The antibacterial activity effect of table 2.A.C.C. extract

Therefore, it is known that A.C.C. extract is the effective component for having potent antibacterial effect to above 5 kinds of bacterium.

Influence of the 5.A.C.C. extract to the diaper rash, prickly heat of infant

Infant with diaper rash, pruritus, prickly heat symptom is faced when being coated with A.C.C. extract as object Confirmed on bed eruption, allergic skin, fester, whether the symptoms such as pruritus, prickly heat are improved.Before clinical test, to go out Rash allergic skin, is festered, 5 kinds of projects in total such as pruritus and prickly heat, and preparatory questionnaire survey is carried out.With 4 points in total for full marks It carries out, eruption project is 2.61 ± 1.1 points, highest, and secondly the high prickly heat project of score is 2.41 ± 1.1 points.Production is asked in advance After volume, daily in the same time, one day 5~10 times coating 1cc amounts, so coating 14 days, after spending 14 days, production is tested Questionnaire.As a result, the use of rear score being 1.5 points hereinafter, the form improved on the whole is presented.Use preceding highest eruption It is all respectively 1.39 ± 0.70,1.35 ± 0.70 points with prickly heat, confirmation reduces to obtain considerable result (Fig. 5, table 3).Also, To before the clinical test of infant with by A.C.C. extract be coated with 1 week after, and by A.C.C. extract be coated with 2 weeks it Process afterwards is compared and observes.As a result, can confirm when the prickly heat of skin is passed through 2 weeks and disappear quite a lot of.Therefore, A.C.C. extract is considered as (referring to Fig. 6) effective to the symptom of diaper rash and prickly heat etc..

Table 3

Table 3.A.C.C. extract influences the diaper rash of infant, prickly heat bring

By above-mentioned experimental result of the invention, A.C.C. extract can be confirmed in the RAW as mouse macrophage Do not have cytotoxicity (Fig. 2) in 264.7 cells, it means that the effect that A.C.C. extract is presented in cell is and cell Survival rate the unrelated intrinsic effect of variation.

In order to confirm the anti-inflammatory effect of A.C.C. extract, with various concentration, A.C.C. extract is pre-processed in RAW 264.7 cells 3 hours after handling 1 μ g/ml LPS 24 hours, measure the generation variation of inflammatory factors NO in cell.Its As a result, according to LPS, increased NO production quantity by A.C.C. extract concentrations dependence reduces (Fig. 3).

This result means that A.C.C. extract inhibits important medium factor NO in the inflammatory process carried out by LPS Formation, thus have anti-inflammatory effect.

Also, when starting inflammatory reaction, the transcription factor NF-KB of inflammatory reaction is activated, adjust TNF-α, IL-1 β and The important inflammation of IL-6 etc promotes the gene expression of albumen, it is known that these adjust the activation of iNOS and play weight to inflammatory reaction It acts on.It is thus identified that inflammatory mediator cell factor (TNF-α, IL-1 β and IL-6) band of the important indicator as inflammation The effect of the A.C.C. extract come.LPS is handled in 264.7 cell of RAW, induces inflammation, at this point, confirmation A.C.C. extract How the result of TNF-α, the production quantity of IL-1 β and IL-6 is changed, it is seen that the potent reduction in A.C.C. extract concentrations dependence ground The inflammatory factor (Fig. 4) induced by LPS.

It is clearly learnt by these results, A.C.C. extract reduces the generation of TNF-α, IL-1 β and IL-6, thus presents The anti-inflammatory effect that expression by adjusting iNOS inhibits NO to generate.Also, A.C.C. extract is to Pseudomonas aeruginosa、Staphylococcus aureus、MRSA(Methicillin-resistant Staphylococcus Aureus), Candida albicans and Streptococcus murans bacterium, are presented 99.9% potent bacterial reduction (%) (table 1).This illustrates that A.C.C. extract is the effective inflammation that the synergy of anti-inflammatory effect and antibacterial action is presented Therapeutic agent has utilizability.In order to clearly confirm these, the infant of diaper rash, pruritus, prickly heat symptom will be suffered from As object, whether have when having rated coating A.C.C. eruption, allergic skin, fester, the symptom of pruritus, prickly heat etc. improves Effect.Evaluate preparatory questionnaire the results show that eruption project is 2.61 ± 1.1 points, and highest makes in the case where 4 points is full marks After 14 days, score is 1.39 ± 0.70 points, than reducing quite a lot of before using (Fig. 5).Also, observe facing for infant The result that bed tests forward and backward process can confirm that when using A.C.C. extract 2 weeks, the prickly heat of skin is alleviated quite not It is wrong.This is clearly stated, and A.C.C. extract is clinically also to alleviate diaper rash and prickly heat by anti-inflammatory and antibacterial action Deng symptom substance (Fig. 6).Therefore, there is the A.C.C. extract of anti-inflammatory and antibacterial effect and as main material Product can be used as to the effective alternative solution of diseases associated with inflammation.

Claims

1. a kind of A.C.C. extract, which is characterized in that by Cortex Phellodendri, radix scutellariae, Chinese herbaceous peony, shaggy-fruited dittany, rhizoma anemarrhenae, alum, kapur, Japan Peppermint, elecampane, Korea open country cloves, C. incisa, Eclipta prostrata, honeysuckle and traditional Chinese medicine of licorice root material cooperate.

2. A.C.C. extract according to claim 1, which is characterized in that the cooperation of the Chinese medicine is respectively Cortex Phellodendri 16 ~24 weight percent, 5~11 weight percent of Chinese herbaceous peony, 5~11 weight percent of shaggy-fruited dittany, are known 1~7 weight percent of radix scutellariae Female 1~7 weight percent, 4~12 weight percent of alum, 1~7 weight percent of kapur, 1~7 weight hundred of cornmint Divide ratio, 1~7 weight percent of elecampane, 1~7 weight percent of Korea open country cloves, 1~7 weight percent of C. incisa, murrel 1~7 weight percent of intestines, 1~7 weight percent of honeysuckle, 1~7 weight percent of Radix Glycyrrhizae.

3. A.C.C. extract according to claim 1, which is characterized in that have antibacterial and anti-inflammatory activity.

4. A.C.C. extract according to claim 1, which is characterized in that for Pseudomonas aeruginosa, staphylococcus aureus, Methicillin-resistant staphylococcus aureus, candida albicans and Streptococcus mutans have antibacterial action.

5. A.C.C. extract according to claim 1, which is characterized in that reduce the tumour as pro-inflammatory cytokine Necrosis factor-alpha (tumor necrosis factor- α, TNF-α), interleukin-1 ' beta ' (interleukin-1 β, IL-1 β) And the generation of interleukin-6 (IL-6), there is anti-inflammatory effect.

6. a kind of antibacterial and anti-inflammatory composition, which is characterized in that include A.C.C. described in any one of claims 1 to 5 Extract is as effective component.